Automated Procedure for Measuring Antigenicity of Extracted and Intact Influenza Virus

Abstract

An automated serum-blocking (S-B) technique was developed in an attempt to find an in vitro test for the determination of the antigenicity of extracted influenza vaccines. The S-B test depends on the ability of an antigen to combine with (or block) specific (in this case, hemagglutination-inhibiting) antibodies. After mixing the test virus with a constant amount of specific antiserum and hemagglutinating virus in an Auto Analyzer, chicken erythrocytes were pumped into the system and the mixture was incubated by passing through coils. The hemagglutinated cells were removed and the residual cells were lysed. The optical density was read and recorded automatically. The S-B test was much more reproducible than the chicken cell agglutination (CCA) test. There was good correlation between the S-B and CCA titers of intact influenza virus, but not of ether-extracted influenza virus. The CCA titer of influenza strains of type A was reduced significantly during ether-extraction. The S-B titers indicated that there was no significant loss in specific antigenicity when influenza strains of types A and B were extracted with ether and Tween-80 according to the described procedure. The S-B test seemed to be a true measurement of the total antigens present in influenza vaccines.

Enhanced Immunogenicity in Mice of a Purified, Tween-Ether-Treated Influenza Vaccine

“…Hemagglutination readings were done on intact vaccines by using a Coleman Junior Spectrophotometer (model 6C) at 542 nm. Since the chicken cell agglutination (CCA) test has questionable value with ether-treated influenza virus preparation (4), the HA and ET antigens were standardized by dilution to the same degree as the starting intact preparations (1,11).…”